Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints
Thousands of antibodies for diagnostic and other analytical purposes are on the market. However, it is often difficult to identify duplicates, reagent changes, and to assign the correct original publications to an antibody. This slows down scientific progress and might even be a cause of irreproduci...
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doaj-415a954a87ff4231a710328ff3fbbeac2020-11-25T01:28:24ZengMDPI AGAntibodies2073-44682020-03-0192810.3390/antib9020008antib9020008Fast Confirmation of Antibody Identity by MALDI-TOF MS FingerprintsGeorg Tscheuschner0Timm Schwaar1Michael G. Weller2Federal Institute for Materials Research and Testing (BAM), Division 1.5 Protein Analysis, Richard-Willstätter-Strasse 11, 12489 Berlin, GermanyFederal Institute for Materials Research and Testing (BAM), Division 1.5 Protein Analysis, Richard-Willstätter-Strasse 11, 12489 Berlin, GermanyFederal Institute for Materials Research and Testing (BAM), Division 1.5 Protein Analysis, Richard-Willstätter-Strasse 11, 12489 Berlin, GermanyThousands of antibodies for diagnostic and other analytical purposes are on the market. However, it is often difficult to identify duplicates, reagent changes, and to assign the correct original publications to an antibody. This slows down scientific progress and might even be a cause of irreproducible research and a waste of resources. Recently, activities were started to suggest the sole use of recombinant antibodies in combination with the open communication of their sequence. In this case, such uncertainties should be eliminated. Unfortunately, this approach seems to be rather a long-term vision since the development and manufacturing of recombinant antibodies remain quite expensive in the foreseeable future. Nearly all commercial antibody suppliers also may be reluctant to publish the sequence of their antibodies, since they fear counterfeiting. De novo sequencing of antibodies is also not feasible today for a reagent user without access to the hybridoma clone. Nevertheless, it seems to be crucial for any scientist to have the opportunity to identify an antibody undoubtedly to guarantee the traceability of any research activity using antibodies from a third party as a tool. For this purpose, we developed a method for the identification of antibodies based on a MALDI-TOF MS fingerprint. To circumvent lengthy denaturation, reduction, alkylation, and enzymatic digestion steps, the fragmentation was performed with a simple formic acid hydrolysis step. Eighty-nine unknown monoclonal antibodies were used for this study to examine the feasibility of this approach. Although the molecular assignment of peaks was rarely possible, antibodies could be easily recognized in a blinded test, simply from their mass-spectral fingerprint. A general protocol is given, which could be used without any optimization to generate fingerprints for a database. We want to propose that, in most scientific projects relying critically on antibody reagents, such a fingerprint should be established to prove and document the identity of the used antibodies, as well as to assign a specific reagent to a datasheet of a commercial supplier, public database record, or antibody ID.https://www.mdpi.com/2073-4468/9/2/8antibody idantibody registryresearch resource identifierrridreproducibilityquality controldocumentationtraceabilityclonesbiochemical reagentsdiagnosticsimmunoassayselisawestern blotimmunohistochemistrymicroarraybiosensor |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Georg Tscheuschner Timm Schwaar Michael G. Weller |
spellingShingle |
Georg Tscheuschner Timm Schwaar Michael G. Weller Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints Antibodies antibody id antibody registry research resource identifier rrid reproducibility quality control documentation traceability clones biochemical reagents diagnostics immunoassays elisa western blot immunohistochemistry microarray biosensor |
author_facet |
Georg Tscheuschner Timm Schwaar Michael G. Weller |
author_sort |
Georg Tscheuschner |
title |
Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints |
title_short |
Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints |
title_full |
Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints |
title_fullStr |
Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints |
title_full_unstemmed |
Fast Confirmation of Antibody Identity by MALDI-TOF MS Fingerprints |
title_sort |
fast confirmation of antibody identity by maldi-tof ms fingerprints |
publisher |
MDPI AG |
series |
Antibodies |
issn |
2073-4468 |
publishDate |
2020-03-01 |
description |
Thousands of antibodies for diagnostic and other analytical purposes are on the market. However, it is often difficult to identify duplicates, reagent changes, and to assign the correct original publications to an antibody. This slows down scientific progress and might even be a cause of irreproducible research and a waste of resources. Recently, activities were started to suggest the sole use of recombinant antibodies in combination with the open communication of their sequence. In this case, such uncertainties should be eliminated. Unfortunately, this approach seems to be rather a long-term vision since the development and manufacturing of recombinant antibodies remain quite expensive in the foreseeable future. Nearly all commercial antibody suppliers also may be reluctant to publish the sequence of their antibodies, since they fear counterfeiting. De novo sequencing of antibodies is also not feasible today for a reagent user without access to the hybridoma clone. Nevertheless, it seems to be crucial for any scientist to have the opportunity to identify an antibody undoubtedly to guarantee the traceability of any research activity using antibodies from a third party as a tool. For this purpose, we developed a method for the identification of antibodies based on a MALDI-TOF MS fingerprint. To circumvent lengthy denaturation, reduction, alkylation, and enzymatic digestion steps, the fragmentation was performed with a simple formic acid hydrolysis step. Eighty-nine unknown monoclonal antibodies were used for this study to examine the feasibility of this approach. Although the molecular assignment of peaks was rarely possible, antibodies could be easily recognized in a blinded test, simply from their mass-spectral fingerprint. A general protocol is given, which could be used without any optimization to generate fingerprints for a database. We want to propose that, in most scientific projects relying critically on antibody reagents, such a fingerprint should be established to prove and document the identity of the used antibodies, as well as to assign a specific reagent to a datasheet of a commercial supplier, public database record, or antibody ID. |
topic |
antibody id antibody registry research resource identifier rrid reproducibility quality control documentation traceability clones biochemical reagents diagnostics immunoassays elisa western blot immunohistochemistry microarray biosensor |
url |
https://www.mdpi.com/2073-4468/9/2/8 |
work_keys_str_mv |
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